Method of distinguishing mesenchymal stem cells and method of determining purity of mesenchymal stem cells

A quality stem cell, purity technology, applied in the determination/inspection of microorganisms, embryonic cells, animal cells, etc., can solve problems such as non-existence

Inactive Publication Date: 2018-03-13
MERIDIGEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Currently, there is no accepted standard or single cell surface marker for the isolation of MSCs from fibroblasts

Method used

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  • Method of distinguishing mesenchymal stem cells and method of determining purity of mesenchymal stem cells
  • Method of distinguishing mesenchymal stem cells and method of determining purity of mesenchymal stem cells
  • Method of distinguishing mesenchymal stem cells and method of determining purity of mesenchymal stem cells

Examples

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Effect test

example 1

[0046] Example 1: Flow Cytometry Analysis of Mixed Populations of Mesenchymal Stem Cells (MSCs) and Fibroblasts

[0047] In order to prove that CD146 can be used as a biomarker for the separation of placenta-derived mesenchymal stem cells (MSCs) and fibroblasts, MSCs and fibroblasts derived from umbilical cord were mixed at the following ratio (MSC : number of cells of fibroblasts) mix: 2x10 5 : 0, 1.8x10 5 : 2x10 4 , 1x10 5 : 1x10 5 , 2x10 4 : 1.8x10 5 , or 0: 2x10 5 . CD146 in each microcentrifuge tube was then analyzed by flow cytometry + group. refer to figure 1 , the results showed that through flow cytometry, CD146 detected by anti-CD146 antibody (anti-human CD146-V450, BD biosciences company) + The percentage of the population decreases proportionally to the increasing fibroblast population.

example 2

[0048] Example 2: The expression level of CD146 is still maintained as the number of subcultures increases

[0049] The MSCs derived from the umbilical cord were harvested at different passage numbers (passage numbers: 4, 6, 8, 10, and 12), and the expression of CD146 was analyzed by flow cytometry. The results showed that after subculture, the expression level of CD146 was still maintained ( figure 2 ).

example 3

[0050] Example 3: Assessment of the purity of MSCs

[0051] In a microcentrifuge tube, mix umbilical cord-derived MSCs with fibroblasts in the following ratio (MSC: cell number of fibroblasts): 2x10 5 : 0, 1.8x10 5 : 2x10 4 , 1x10 5 : 1x10 5 , 2x10 4 : 1.8x10 5 , or 0: 2x10 5 . CD146 in each microcentrifuge tube was then analyzed by flow cytometry + group. Subsequently, the relationship between the purity (%) of MSCs and the percentage of cells expressing CD146 was analyzed by simple linear regression, the results of which are shown in image 3 . The regression equation is Y=1.206X-23.32, where Y is the purity (%) of MSCs, and X is the percentage of cells expressing CD146. The results confirmed that there is a linear relationship between the expression level of CD146 and the purity of MSC.

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Abstract

A method of distinguishing mesenchymal stem cells (MSCs) from fibroblasts is provided. Also provided is a method of increasing a purity of mesenchymal stem cells (MSCs) population in a cell culture. The above-mentioned methods each comprise a step of sorting or isolating the cells by a marker of CD146 from a cell culture derived from a placenta-related tissue. Further provided is a method of assessing purity of mesenchymal stem cells (MSCs) in a cell culture derived from a placenta-related tissue, comprising determining the percentage of cells expressing CD146 in the culture.

Description

technical field [0001] The present invention relates to a method of distinguishing mesenchymal stem cells (MSCs) in cell culture derived from placenta-associated tissue. The present invention also relates to methods of increasing the purity of a population of mesenchymal stem cells (MSCs) in cell culture derived from placenta-associated tissue. In another aspect, the present invention relates to a method of assessing the purity of mesenchymal stem cells (MSCs) in cell culture derived from placenta-associated tissue. Background technique [0002] Mesenchymal stem cells or stromal cells (Mesenchymal stem or stromal cells, MSCs) are pluripotent cells derived from the embryonic mesoderm and have a fibroblast-like morphology. The cells can be differentiated into fat cells, bone cells, chondrocytes, nerve cells, muscle cells and other cell types depending on the stimulation and culture conditions. Although the plasticity of human mesenchymal stem cells (hMSCs) and their role in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N15/14
CPCG01N15/14G01N33/53C12N5/0605C12N5/0668C12N2501/599G01N33/56966G01N2333/70596C12N5/0662G01N35/0098G01N2015/1488C12Q1/6881C12Q2600/158
Inventor 宣昶有林卫理苏郁清刘威廷李孟玮杜茂宽
Owner MERIDIGEN BIOTECH
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